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. 2008 Jul 1;64(Pt 7):629-31.
doi: 10.1107/S1744309108016059. Epub 2008 Jun 11.

Crystallization and preliminary X-ray characterization of the genetically encoded fluorescent calcium indicator protein GCaMP2

María M Rodríguez Guilbe  1 , Elisa C Alfaro Malavé, Jasper Akerboom, Jonathan S Marvin, Loren L Looger, Eric R Schreiter
Affiliations

Crystallization and preliminary X-ray characterization of the genetically encoded fluorescent calcium indicator protein GCaMP2

María M Rodríguez Guilbe et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

Abstract

Fluorescent proteins and their engineered variants have played an important role in the study of biology. The genetically encoded calcium-indicator protein GCaMP2 comprises a circularly permuted fluorescent protein coupled to the calcium-binding protein calmodulin and a calmodulin target peptide, M13, derived from the intracellular calmodulin target myosin light-chain kinase and has been used to image calcium transients in vivo. To aid rational efforts to engineer improved variants of GCaMP2, this protein was crystallized in the calcium-saturated form. X-ray diffraction data were collected to 2.0 A resolution. The crystals belong to space group C2, with unit-cell parameters a = 126.1, b = 47.1, c = 68.8 A, beta = 100.5 degrees and one GCaMP2 molecule in the asymmetric unit. The structure was phased by molecular replacement and refinement is currently under way.

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Figures

Figure 1
Figure 1
SDS–PAGE analysis of the purified GCaMP2 protein used for crystallization. Molecular weights of protein markers (in kDa) are indicated on the left.
Figure 2
Figure 2
Crystals of calcium-saturated GCaMP2. The scale bar is 100 µm in length.
Figure 3
Figure 3
Representative X-ray diffraction pattern from a single GCaMP2 crystal. The smaller and larger circles correspond to Bragg spacings of 3.0 and 2.0 Å, respectively.
See this image and copyright information in PMC

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